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Early Stages of Golgi Vesicle and Tubule Formation Require Diacylglycerol

2008; American Society for Cell Biology; Volume: 20; Issue: 3 Linguagem: Inglês

10.1091/mbc.e08-03-0256

1939-4586

Lennart Asp, Fredrik Kartberg, Julia Fernández-Rodrı́guez, Maria Smedh, Markus Elsner, Fréderic Laporte, Montserrat Bárcena, Karen A. Jansen, Jack A. Valentijn, Abraham J. Koster, John Bergeron, Tommy Nilsson,

Endoplasmic Reticulum Stress and Disease

We have investigated the role for diacylglycerol (DAG) in membrane bud formation in the Golgi apparatus. Addition of propranolol to specifically inhibit phosphatidate phosphohydrolase (PAP), an enzyme responsible for converting phosphatidic acid into DAG, effectively prevents formation of membrane buds. The effect of PAP inhibition on Golgi membranes is rapid and occurs within 3 min. Removal of the PAP inhibitor then results in a rapid burst of buds, vesicles, and tubules that peaks within 2 min. The inability to form buds in the presence of propranolol does not appear to be correlated with a loss of ARFGAP1 from Golgi membranes, as knockdown of ARFGAP1 by RNA interference has little or no effect on actual bud formation. Rather, knockdown of ARFGAP1 results in an increase in membrane buds and a decrease of vesicles and tubules suggesting it functions in the late stages of scission. How DAG promotes bud formation is discussed.